1. Simultaneous Spectrophotometric Quantitation of Cu(II) and Ni(II)
Lab 8

2. Outline Purpose Absorbance – Single Analyte
Absorbance – Multiple Analyte
Solving unknown concentrations
Procedure
Safety Concerns
Waste
Next Lab Reminder

3. Purpose
The purpose of this lab is to demonstrate the additive property of absorbance. The molar absorptivity (ε) values for both Cu(II) and Ni(II) will be found at two analytical wavelengths by measuring the absorbance of both ions in solutions of known concentration. The two analytical wavelengths will then be utilized to find the concentrations of both Cu(II) and Ni(II) in an unknown mixture.

4. Absorbance – Beer’s Law
For a single analyte, we can use the Beer-Lambert Law:
Abs = ε b c to determine any one of the variables, given that three of them are already known.
Since molar absorptivity (ε ) is the same for a given compound (x), regardless of concentration, at a given wavelength, we can clarify Beer’s Law terms:
Aλ1 = ελ1x b [x]
Given that the path length (b) through our cuvets is cm, we can define a new variable kλ1x = ελ1x b to simplify calculations and modify Beer’s Law accordingly: Aλ1 = kλ1x [x]
After you make up your solutions today, you will simply use this modified Beer’s Law to determine the molar absorptivity (kλ1x ) of each solution at a given wavelength. The average k value for each set of solutions at each wavelength is calculated and used to solve for unknown concentrations.

5. Absorbance is Additive
For multiple analytes we exploit the additive property of absorbance: AbsT = Abs1 + Abs
The same number of wavelengths are used as number of analytes that are analyzed.
Therefore, in two analytes, we use:
Aλ1 = ελ1x b [x] + ελ1y b [y];
Define kλ1x = ελ1x b and kλ1y = ελ1y b;
So Aλ1 = kλ1x [x] + kλ1y [y]
Aλ2 = ελ2x b [x] + ελ2y b [y];
Define kλ2x = ελ2x b and kλ2y = ελ2y b;
So Aλ2 = kλ2x [x] + kλ2y [y]

6. At 395 nm, only Ni2+ absorbs At 395 nm, this equation becomes:
Beer’s Law states:
Aλ1 = kλ1x [x] + kλ1y [y]
At 395 nm, this equation becomes:
A395 = k395Ni(II)[Ni2+] + k395Cu(II)[Cu2+] = k395 [Ni2+] + 0
To solve for [Ni2+] in our unknown solution:
[Ni2+] =

7. At 775 nm both Ni2+ and Cu2+ absorb
Beer’s Law states:
Aλ2 = kλ2x [x] + kλ2y [y]
To solve for [y], [y] =
To solve for [Cu2+] in our unknown solution:
[Cu(II)] =

8. Procedure Make up 4 solutions of Cu2+(aq) and 4 solutions of Ni2+(aq).
Determine the k value of each solution at the indicated wavelengths.
Find the average k values for Cu2+(aq) at 775 nm and for Ni2+(aq) at 775 nm and 395 nm.
Solve for the unknown concentrations of your unknown solution using the derived equations.

9. Safety Concerns Reagents: Eye Contact: Skin Contact: Inhalation:
Cupric Sulfate
Nickel Sulfate / Nickel Chloride
Eye Contact:
Irritation, redness, pain, conjunctivitis, ulceration, clouding of cornea
Skin Contact:
Irritation, redness, rash and itching. Sensitizer.
Inhalation:
Coughing, sore throat, shortness of breath, ulceration and perforation of the respiratory tract. Fumes from heating may cause symptoms similar to a cold. May cause metallic taste in mouth. Lung damage, allergy and asthma may occur.
Ingestion:
Burning of the mouth, esophagus, and stomach. Hemorrhagic gastritis, nausea, vomiting, abdominal pain, giddiness, myocardial weakness, metallic taste, and diarrhea. Systemic copper poisoning with capillary damage, headache, cold sweat, weak pulse, kidney and liver damage, CNS excitation and depression, jaundice, convulsions, blood effects, paralysis, coma and death.

10. Waste Copper and Nickel are both toxic.
Dispose of them in the appropriate container(s) in the fume hood.

11. Lab 9 Reminder
Lab 9 is next.